Objective:To construct four recombinant hemagglutinin (HA) antigens from seasonal influenza viruses and evaluate their immunogenicity in mouse models.Methods:HA coding sequences from four seasonal influenza virus strains Wisconsin (H1N1), Darwin (H3N2), Austria (B/Victoria lineage, BV) and Phuket (B/Yamagata lineage, BY) were optimized and synthesized, and then used to construct four recombinant plasmids. Recombinant baculoviruses were obtained through transformation and transfection. The expression of recombinant HA antigens was identified by SDS-PAGE and Western blot. The recombinant HA antigens were purified by nickel column affinity chromatography and intramuscularly administered to BALB/c mice after formulation with Al(OH) 3 or AddaVax adjuvant. Humoral immune responses were assessed by indirect ELISA and hemagglutination inhibition test, while cellular immune responses were evaluated by ELISPOT. Microneutralization test was used to detect the titers of serum antibodies in mice. Statistical analysis was performed using t test or non-parametric rank sum test. Results:PCR amplification and agarose gel electrophoresis confirmed the correct construction of the recombinant bacmids. Western blot showed verified the successful expression of the four recombinant antigens (H1-HA, H3N2-HA, BV-HA, and BY-HA). SDS-PAGE results showed that the purity of all four recombinant HA antigens exceeded 95%. After three-dose immunization, the total IgG levels in mice immunized with the recombinant H1N1-HA, H3N2-HA, or BV-HA formulated with AddaVax adjuvant were higher than those in the corresponding groups immunized with the same recombinant antigen alone (all P<0.05). The secretion levels of IFN-γ, IL-2, and IL-4 in the group receiving the mixture of all four recombinant HA antigens formulated with AddaVax adjuvant were higher than those in the group immunized with a commercial quadrivalent split influenza vaccine (all P<0.01). Results of the microneutralization test showed that the antibody titer in the quadrivalent split influenza vaccine group was 1∶225, whereas the titer in the group immunized with the mixture of four recombinant HA antigens formulated with AddaVax adjuvant could reach up to 1∶1 200. Conclusions:In this study, four recombinant seasonal influenza virus HA antigens are successfully expressed and demonstrated good immunogenicity in mice when formulated AddaVax adjuvant.

Objective:To construct four recombinant hemagglutinin (HA) antigens from seasonal influenza viruses and evaluate their immunogenicity in mouse models.Methods:HA coding sequences from four seasonal influenza virus strains Wisconsin (H1N1), Darwin (H3N2), Austria (B/Victoria lineage, BV) and Phuket (B/Yamagata lineage, BY) were optimized and synthesized, and then used to construct four recombinant plasmids. Recombinant baculoviruses were obtained through transformation and transfection. The expression of recombinant HA antigens was identified by SDS-PAGE and Western blot. The recombinant HA antigens were purified by nickel column affinity chromatography and intramuscularly administered to BALB/c mice after formulation with Al(OH) 3 or AddaVax adjuvant. Humoral immune responses were assessed by indirect ELISA and hemagglutination inhibition test, while cellular immune responses were evaluated by ELISPOT. Microneutralization test was used to detect the titers of serum antibodies in mice. Statistical analysis was performed using t test or non-parametric rank sum test. Results:PCR amplification and agarose gel electrophoresis confirmed the correct construction of the recombinant bacmids. Western blot showed verified the successful expression of the four recombinant antigens (H1-HA, H3N2-HA, BV-HA, and BY-HA). SDS-PAGE results showed that the purity of all four recombinant HA antigens exceeded 95%. After three-dose immunization, the total IgG levels in mice immunized with the recombinant H1N1-HA, H3N2-HA, or BV-HA formulated with AddaVax adjuvant were higher than those in the corresponding groups immunized with the same recombinant antigen alone (all P<0.05). The secretion levels of IFN-γ, IL-2, and IL-4 in the group receiving the mixture of all four recombinant HA antigens formulated with AddaVax adjuvant were higher than those in the group immunized with a commercial quadrivalent split influenza vaccine (all P<0.01). Results of the microneutralization test showed that the antibody titer in the quadrivalent split influenza vaccine group was 1∶225, whereas the titer in the group immunized with the mixture of four recombinant HA antigens formulated with AddaVax adjuvant could reach up to 1∶1 200. Conclusions:In this study, four recombinant seasonal influenza virus HA antigens are successfully expressed and demonstrated good immunogenicity in mice when formulated AddaVax adjuvant.

Background::Ainuovirine (ANV) is a new generation of non-nucleoside reverse transcriptase inhibitor for the treatment of human immunodeficiency virus (HIV) type 1 infection. This study aimed to evaluate the population pharmacokinetic (PopPK) profile and exposure-response relationship of ANV among people living with HIV.Methods::Plasma concentration-time data from phase 1 and phase 3 clinical trials of ANV were pooled for developing the PopPK model. Exposure estimates obtained from the final model were used in exposure-response analysis for virologic responses and safety responses.Results::ANV exhibited a nonlinear pharmacokinetic profile, which was best described by a two-compartment model with first-order elimination. There were no significant covariates correlated to the pharmacokinetic parameters of ANV. The PopPK parameter estimate (relative standard error [%]) for clearance adjusted for bioavailability (CL/F) was 6.46 (15.00) L/h, and the clearance of ANV increased after multiple doses. The exposure-response model revealed no significant correlation between the virologic response (HIV-RNA <50 copies/mL) at 48 weeks and the exposure, but the incidence of adverse events increased with the increasing exposure ( P value of steady-state trough concentration and area under the steady-state curve were 0.0177 and 0.0141, respectively). Conclusions::Our PopPK model supported ANV 150 mg once daily as the recommended dose for people living with HIV, requiring no dose adjustment for the studied factors. Optimization of ANV dose may be warranted in clinical practice due to an increasing trend in adverse reactions with increasing exposure.Trial registration::Chinese Clinical Trial Registry https://www.chictr.org.cn (Nos. ChiCTR1800018022 and ChiCTR1800019041).

Production internship is an important teaching tache for undergraduate students to carry out engineering training by using professional skills, and it is a key starting point for fostering application-oriented talents in biotechnology. The Course Group of 'production internship of biotechnology majors' of Binzhou University is investigating application-oriented transformation for local regular colleges and universities, as well as fostering high-level application-oriented talents. By taking green fluorescent protein (GFP) polyclonal antibody as an example, the reform and practice on teaching content, teaching mode, assessment method, continuous improvement of curriculum were carried out. Moreover, the characteristics of the Yellow River Delta-Binzhou Biotechnology & Pharmaceutical Industrial Cluster were taken into account to intensify academic-enterprise cooperation. On one hand, this Course Group designed and rearranged the course contents, carried out essential training through online resources and platforms such as virtual simulation, and recorded, tracked and monitored the progress of production internship through practical testing and software platforms like 'Alumni State'. On the other hand, this Course Group established a practice-and application-oriented assessment method in the process of production internship and a dual evaluation model for continuous improvement. These reform and practices have promoted the training of application-oriented talents in biotechnology, and may serve as a reference for similar courses.
Humans ; Internship and Residency ; Curriculum ; Students ; Biotechnology

OBJECTIVE: To analyze the molecular characteristics of a ABO subgroup. METHODS: The ABO phenotype was determined with the tube method. Exons of the ABO gene were analyzed by Sanger sequencing, and haplotypes of exons 6 and 7 were analyzed by cloning sequencing. RESULTS: By forward typing, the red blood cells showed 3+ agglutination reaction with anti-A and 4+ agglutination with anti-B. A weak reaction with A1 cells and no agglutination reaction with B, O cells by the reverse typing. Sequencing results showed heterozygosity including c.297A>G, c.467C>T, c.526C>G, c.608A>G, c.657C>T, c.703G>A, c.796C>A, c.803G>C, c.930G>A. Cloning sequencing revealed a c.608A>G variant in the A allele compared with the ABO*A1.02. CONCLUSION A new variant site of subtype A of c.608G variation has been identified.
ABO Blood-Group System/genetics* ; Alleles ; Exons ; Genotype ; Heterozygote ; Phenotype

Objective:To establish the Sunitinib-resistant model GIST882-Rs.Methods:The intermittent concentration gradient doubling method was used to get Sunitinib-resistant cell line GIST 882-Rs. Changes of cell morphology were observed and compared under the opticalm icroscopy. Changes of cell cycle were assayed and compared by using flow cytometer. The growth curvature , half maximal inhibitory concentration (IC50) were calculated by using cell counting kit (CCK8) assay.Results:Significant changes occurred in the morphologic profiles, such as epithelioid shape, the increased ratio of nucleolus to cytoplasm, emerging multinucleated cel1 and increase of granule sub stances. IC50 for GIST882 and GIST 882-Rs were(127.50 ± 2.15) μmol and (23.98 ± 1.85) μmol, and there were significant differences ( P < 0.05). The resistance index of GIST882-Rs to Sunitinib was 5.30 fold of GIST882 cells. The cell cycle of GIST882 and GIST882- Rs was respectively 52.51% and 39.87% in S phase 25.22% and 32.32% in G 0/G 1 phase. Conclusions:The sensitivity of GIST 882-Rs to Sunitinib treatment is decreased significantly.

AIM:To investigate the serum miRNA-101 expression level in patients with newly diagnosed type 2 diabetes mellitus (T2DM),and to evaluate the clinical implications of miRNA-101 expression level variation.METHODS:qRT-PCR was used to determine the serum miRNA-101 expression level.Pearson correlation analysis was performed to observe the relationship between two variables.Multiple stepwise linear regression analysis was used to assess the association of serum miRNA-101 level and other parameters.RESULTS:Serum miRNA-101 level in patients with newly diagnosed T2DM was significantly higher than that in control subjects (P ＜ 0.05).The serum level of miRNA-101 was positively correlated with the glycosylated hemoglobin A1c (HbA1c,P ＜0.05).Multiple linear regression analysis revealed that the circulating miRNA-101 was in significant positive correlation with HbA1c (P ＜ 0.05) after adjustment for age,sex and body weight.CONCLUSION:Enhanced circulating miRNA-101 level in newly diagnosed T2DM patients may be associated with elevation of HbA1 c.

BACKGROUND/AIMS: The past decades have seen increasing incidence and prevalence of inflammatory bowel disease (IBD) in China. This article aimed to summarize the current status and characteristics of surgical management for IBD in China. METHODS: We searched PubMed, Embase, and Chinese databases from January 1, 1990 to July 1, 2014 for all relevant studies on the surgical treatment IBD in China. Eligible studies with sufficient defined variables were further reviewed for primary and secondary outcome measures. RESULTS: A total of 74 studies comprising 2,007 subjects with Crohn's disease (CD) and 1,085 subjects with ulcerative colitis (UC) were included. The percentage of CD patients misdiagnosed before surgery, including misdiagnosis as appendicitis or UC, was 50.8%±30.9% (578/1,268). The overall postoperative complication rate was 22.3%±13.0% (267/1,501). For studies of UC, the overall postoperative complication rate was 22.2%±27.9% (176/725). In large research centers (n>50 surgical cases), the rates of emergency operations for CD (P=0.032) and in-hospital mortalities resulting from both CD and UC were much lower than those in smaller research centers (n≤50 surgical cases) (P=0.026 and P <0.001, respectively). Regarding the changes in CD and UC surgery over time, postoperative complications (P=0.045 for CD; P=0.020 for UC) and postoperative in-hospital mortality (P=0.0002 for CD; P=0.0160 for UC) both significantly improved after the year 2010. CONCLUSIONS: The surgical management of IBD in China has improved over time. However, the rates of misdiagnosis and postoperative complications over the past two decades have remained high. Large research centers were found to have relatively better capacity for surgical management than the smaller ones. Higher quality prospective studies are needed in China.
Appendicitis ; Asian Continental Ancestry Group ; China* ; Colitis, Ulcerative ; Crohn Disease ; Diagnostic Errors ; Emergencies ; Hospital Mortality ; Humans ; Incidence ; Inflammatory Bowel Diseases* ; Outcome Assessment (Health Care) ; Postoperative Complications ; Prevalence ; Prospective Studies

BACKGROUND/AIMS: The past decades have seen increasing incidence and prevalence of inflammatory bowel disease (IBD) in China. This article aimed to summarize the current status and characteristics of surgical management for IBD in China. METHODS: We searched PubMed, Embase, and Chinese databases from January 1, 1990 to July 1, 2014 for all relevant studies on the surgical treatment IBD in China. Eligible studies with sufficient defined variables were further reviewed for primary and secondary outcome measures. RESULTS: A total of 74 studies comprising 2,007 subjects with Crohn's disease (CD) and 1,085 subjects with ulcerative colitis (UC) were included. The percentage of CD patients misdiagnosed before surgery, including misdiagnosis as appendicitis or UC, was 50.8%±30.9% (578/1,268). The overall postoperative complication rate was 22.3%±13.0% (267/1,501). For studies of UC, the overall postoperative complication rate was 22.2%±27.9% (176/725). In large research centers (n>50 surgical cases), the rates of emergency operations for CD (P=0.032) and in-hospital mortalities resulting from both CD and UC were much lower than those in smaller research centers (n≤50 surgical cases) (P=0.026 and P <0.001, respectively). Regarding the changes in CD and UC surgery over time, postoperative complications (P=0.045 for CD; P=0.020 for UC) and postoperative in-hospital mortality (P=0.0002 for CD; P=0.0160 for UC) both significantly improved after the year 2010. CONCLUSIONS: The surgical management of IBD in China has improved over time. However, the rates of misdiagnosis and postoperative complications over the past two decades have remained high. Large research centers were found to have relatively better capacity for surgical management than the smaller ones. Higher quality prospective studies are needed in China.
Appendicitis ; Asian Continental Ancestry Group ; China* ; Colitis, Ulcerative ; Crohn Disease ; Diagnostic Errors ; Emergencies ; Hospital Mortality ; Humans ; Incidence ; Inflammatory Bowel Diseases* ; Outcome Assessment (Health Care) ; Postoperative Complications ; Prevalence ; Prospective Studies

Objective To evaluate long-term outcome of simultaneous kissing sirolimus-eluting stent (SKS) technique versus single sirolimus-eluting stent (SSS) technique for percutaneous treatment of true coronary bifurcation lesions in large-size vessels.Methods This randomized study assigned 190 patients with a coronary bifurcation lesion to simultaneous kissing stenting (SKS) in both main and side branches and 190 patients to main vessel stenting only (SSS).The endpoints included restenosis,death,non-fatal myocardial infarction,target-lesion revascularization (TLR),stent thrombosis,success rate of percutaneous coronary intervention (PCI) and the operation duration.Results During 1-year follow-up,the SKS group and the SSS group had similar incidences of overall re stenosis [30 ( 15.8 ％ ) vs.24 ( 15.2 ％ ),x2=0.000,P＜0.05],mainbranch restenosis [20 ( 10.5％ ) vs.16 ( 10.1％ ),x2=0.003,P ＞ 0.05];side-branch restenosis [13 ( 6.8％ )vs.23 ( 14.6％ );x2=4.73,P＜0.05];death [2 ( 1.1％ ) vs.1 ( 0.6％ ),x2=0.026,P ＞ 0.05],non-fatal myocardial infarction [4 (2.1％ ) vs.2 ( 1.3％ ),x2=0.034,P ＞ 0.05],TLR [23 ( 12.1％ ) vs.20 ( 12.7％ ),x2=0.000,P ＞ 0.05] and stent thrombosis [4 (2.1％ ) vs.2 ( 1.3 ％ ),x2=0.034,P ＞ 0.05] and a shorter operation duration[(20 ± 8) min vs.(45 ± 9) min,t=1.98,P＜0.05] than the SSS group.Conclusion For true coronary bifurcation lesions in large-size vessels,SKS and SSS have similar long-term outcomes.The SKS group has a higher success rate of PCI and shorter operation duration.